YALEPIC^® Universal qPCR Master Mix with UDG (SYBR)

The UDG (Uracil-DNA Glycosylase) contamination prevention system replaces traditional dTTP with dUTP incorporated into DNA. Before PCR, heat-labile UDG enzyme hydrolyzes uracil-containing contaminating DNA at room temperature, preventing it from being amplified. During PCR thermal cycling, UDG is inactivated and does not affect subsequent reactions. This technology effectively prevents false positives caused by laboratory aerosol contamination, particularly important for clinical detection and high-sensitivity gene quantification research.

Antibody-modified hot-start enzymes physically block Taq enzyme active sites through antibodies at room temperature. Upon heating to hot-start temperature (typically >95 degrees C), the antigen-antibody complex denatures and releases enzyme activity. Compared to chemical modification, the antibody method offers faster activation, lower residual activity, and stronger inhibition of non-specific amplification, particularly suitable for high-sensitivity detection of low-copy templates.

High GC templates easily form secondary structures. This product’s buffer has been optimized for high GC systems. If amplification results are still unsatisfactory, recommendations: (1) Optimize annealing temperature (increase by 3-5 degrees C); (2) Add 5-10% DMSO or betaine as final concentration to assist strand separation; (3) Ensure complete template denaturation (extend initial denaturation at 95 degrees C to 5-10 minutes).

It is recommended to test each primer at a final concentration starting from 0.2 microM, with optimization in the 0.1-0.5 microM range. If non-specific amplification or primer-dimers occur (melt curve extra peaks), try reducing primer concentration to 0.1-0.15 microM. If amplification efficiency is low, test in a 0.2-0.4 microM gradient.

Check the following points: (1) Template concentration too high or too low (cDNA template recommended 1-10 ng/reaction, genomic DNA 10-100 ng/reaction); (2) Primer design should avoid consecutive GC at 3′ end; (3) Confirm ROX reference dye type matches instrument (this product’s universal ROX is compatible with all models); (4) Check that the amplification system is thoroughly mixed – SYBR Green I dye should not be vigorously shaken to produce bubbles; (5) Ensure precipitate is fully dissolved and mixed after reagent thawing.

This product may show white precipitate after low-temperature storage or freezing, which is a normal phenomenon of SYBR Green I dye and buffer salt precipitation. Place at room temperature protected from light for 3-5 minutes, gently invert to mix (do not vortex vigorously) until precipitate is fully dissolved. This does not affect reagent performance.

This product contains pre-mixed universal ROX reference dye, compatible with all mainstream qPCR instruments: ABI (7500/7500 Fast/ViiA7/QuantStudio series), Bio-Rad (CFX96/CFX Connect, etc.), Roche LightCycler series, Agilent Mx series, Jena, and other brands. No additional ROX purchase needed, greatly simplifying the experimental workflow.

This product is a SYBR Green dye-based qPCR master mix and does not contain reverse transcriptase components. For one-step qPCR, use with YALI Biotech reverse transcription kits, or select YALI Biotech one-step qPCR master mix product series. YALEPIC U+ One Step qRT-PCR Probe MasterMix CAT#YPQ42003.

For gene expression differential analysis, primer screening, and other rapid quantification, this SYBR Green method (universal high-sensitivity) is recommended. For multiplex detection, absolute quantification, or clinical-grade high-specificity detection, YALI Biotech probe-based qPCR master mix product series CAT#YQ32032 is recommended.

YALI Biotech offers trial samples for the full range of YALEPIC qPCR master mix products for new customers. You can apply through the official customer service hotline (0512-66256010) or email info@yalibiotech.com. Submit your research direction and estimated usage to receive free samples.